Cells are exquisitely compartmentalized to achieve precise spatiotemporal regulation of myriad processes and pathways. Phase separation offers one way to achieve territorial organization in the cellular context, via the creation of membrane-less organelles (MLOs). MLOs formed through phase separation are associated with numerous critical biological functions. Although hundreds of publications on related topics are produced each year, robust criteria for the determination of biologically meaningful phase separation are yet to be well established. Here we present some principles and propose a few guidelines for phase-separation studies in biology. Specifically, we provide an in-depth experiment pipeline for phase-separation studies, including mechanisms of the molecular driving forces, ways to correlate in vivo and in vitro observations, and strategies to relate the phase-separation phenomenon to biological functions. We also intend to contribute to streamlining the aforementioned diagnostic criteria by further stressing a few common caveats in the field.

细胞被巧妙地分隔开，以实现对无数过程和通路的精确时空调节。通过创建无膜细胞器 (MLO)，相分离提供了一种在细胞环境中实现区域组织的方法。通过相分离形成的 MLO 与许多关键的生物学功能相关。尽管每年都会产生数百篇关于相关主题的出版物，但尚未确定确定具有生物学意义的相分离的稳健标准。在这里，我们提出了一些原则，并提出了一些生物学相分离研究的指导方针。具体来说，我们为相分离研究提供了深入的实验流程，包括分子驱动力的机制、体内和体外观察的关联方法、以及将相分离现象与生物功能联系起来的策略。我们还打算通过进一步强调该领域的一些常见警告来帮助简化上述诊断标准。